CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Maarten H Geurts; Eyleen de Poel; Gimano D Amatngalim; Rurika Oka; Fleur M Meijers; Evelien Kruisselbrink; Peter van Mourik; Gitte Berkers; Karin M de Winter-de Groot; Sabine Michel; Danya Muilwijk; Bente L Aalbers; Jasper Mullenders; Sylvia F Boj; Sylvia W F Suen; Jesse E Brunsveld; Hettie M Janssens; Marcus A Mall; Simon Y Graeber; Ruben van Boxtel; Cornelis K van der Ent; Jeffrey M Beekman; Hans Clevers

doi:10.1016/j.stem.2020.01.019

CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Maarten H Geurts, Eyleen de Poel, Gimano D Amatngalim, Rurika Oka, Fleur M Meijers, Evelien Kruisselbrink, Peter van Mourik, Gitte Berkers, Karin M de Winter-de Groot, Sabine Michel, Danya Muilwijk, Bente L Aalbers, Jasper Mullenders, Sylvia F Boj, Sylvia W F Suen, Jesse E Brunsveld, Hettie M Janssens, Marcus A Mall, Simon Y Graeber, Ruben van BoxtelCornelis K van der Ent, Jeffrey M Beekman, Hans Clevers

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

117 Citations (Scopus)

Abstract

Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs. ABE holds promise for clinical application, as it does not depend on the introduction of double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering. Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing 664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE (PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on four selected CF organoid samples. Genetic and functional repair was obtained in all four cases, while whole-genome sequencing (WGS) of corrected lines of two patients did not detect off-target mutations. These observations exemplify the value of large, patient-derived organoid biobanks representing hereditary disease and indicate that ABE may be safely applied in human cells.

Original language	English
Pages (from-to)	503-510.e7
Journal	Cell Stem Cell
Volume	26
Issue number	4
DOIs	https://doi.org/10.1016/j.stem.2020.01.019
Publication status	Published - 02 Apr 2020

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Geurts, M. H., de Poel, E., Amatngalim, G. D., Oka, R., Meijers, F. M., Kruisselbrink, E., van Mourik, P., Berkers, G., de Winter-de Groot, K. M., Michel, S., Muilwijk, D., Aalbers, B. L., Mullenders, J., Boj, S. F., Suen, S. W. F., Brunsveld, J. E., Janssens, H. M., Mall, M. A., Graeber, S. Y., ... Clevers, H. (2020). CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank. Cell Stem Cell, 26(4), 503-510.e7. https://doi.org/10.1016/j.stem.2020.01.019

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title = "CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank",

abstract = "Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs. ABE holds promise for clinical application, as it does not depend on the introduction of double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering. Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing 664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE (PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on four selected CF organoid samples. Genetic and functional repair was obtained in all four cases, while whole-genome sequencing (WGS) of corrected lines of two patients did not detect off-target mutations. These observations exemplify the value of large, patient-derived organoid biobanks representing hereditary disease and indicate that ABE may be safely applied in human cells.",

author = "Geurts, {Maarten H} and {de Poel}, Eyleen and Amatngalim, {Gimano D} and Rurika Oka and Meijers, {Fleur M} and Evelien Kruisselbrink and {van Mourik}, Peter and Gitte Berkers and {de Winter-de Groot}, {Karin M} and Sabine Michel and Danya Muilwijk and Aalbers, {Bente L} and Jasper Mullenders and Boj, {Sylvia F} and Suen, {Sylvia W F} and Brunsveld, {Jesse E} and Janssens, {Hettie M} and Mall, {Marcus A} and Graeber, {Simon Y} and {van Boxtel}, Ruben and {van der Ent}, {Cornelis K} and Beekman, {Jeffrey M} and Hans Clevers",

year = "2020",

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doi = "10.1016/j.stem.2020.01.019",

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Geurts, MH, de Poel, E, Amatngalim, GD, Oka, R, Meijers, FM, Kruisselbrink, E, van Mourik, P, Berkers, G, de Winter-de Groot, KM, Michel, S, Muilwijk, D, Aalbers, BL, Mullenders, J, Boj, SF, Suen, SWF, Brunsveld, JE, Janssens, HM, Mall, MA, Graeber, SY, van Boxtel, R, van der Ent, CK, Beekman, JM & Clevers, H 2020, 'CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank', Cell Stem Cell, vol. 26, no. 4, pp. 503-510.e7. https://doi.org/10.1016/j.stem.2020.01.019

TY - JOUR

T1 - CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

AU - Geurts, Maarten H

AU - de Poel, Eyleen

AU - Amatngalim, Gimano D

AU - Oka, Rurika

AU - Meijers, Fleur M

AU - Kruisselbrink, Evelien

AU - van Mourik, Peter

AU - Berkers, Gitte

AU - de Winter-de Groot, Karin M

AU - Michel, Sabine

AU - Muilwijk, Danya

AU - Aalbers, Bente L

AU - Mullenders, Jasper

AU - Boj, Sylvia F

AU - Suen, Sylvia W F

AU - Brunsveld, Jesse E

AU - Janssens, Hettie M

AU - Mall, Marcus A

AU - Graeber, Simon Y

AU - van Boxtel, Ruben

AU - van der Ent, Cornelis K

AU - Beekman, Jeffrey M

AU - Clevers, Hans

PY - 2020/4/2

Y1 - 2020/4/2

N2 - Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs. ABE holds promise for clinical application, as it does not depend on the introduction of double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering. Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing 664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE (PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on four selected CF organoid samples. Genetic and functional repair was obtained in all four cases, while whole-genome sequencing (WGS) of corrected lines of two patients did not detect off-target mutations. These observations exemplify the value of large, patient-derived organoid biobanks representing hereditary disease and indicate that ABE may be safely applied in human cells.

AB - Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs. ABE holds promise for clinical application, as it does not depend on the introduction of double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering. Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing 664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE (PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on four selected CF organoid samples. Genetic and functional repair was obtained in all four cases, while whole-genome sequencing (WGS) of corrected lines of two patients did not detect off-target mutations. These observations exemplify the value of large, patient-derived organoid biobanks representing hereditary disease and indicate that ABE may be safely applied in human cells.

U2 - 10.1016/j.stem.2020.01.019

DO - 10.1016/j.stem.2020.01.019

M3 - Article

C2 - 32084388

SN - 1934-5909

VL - 26

SP - 503-510.e7

JO - Cell Stem Cell

JF - Cell Stem Cell

IS - 4

ER -

CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Abstract

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